Difference between revisions of "Team:UNITN-Trento/Safety"

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<h2>Safety in iGEM</h2>
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<p>Please visit <a href="https://2015.igem.org/Safety">the main Safety page</a> to find this year's safety requirements & deadlines, and to learn about safe & responsible research in iGEM.</p>
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<p>On this page of your wiki, you should write about how you are addressing any safety issues in your project. The wiki is a place where you can <strong>go beyond the questions on the safety forms</strong>, and write about whatever safety topics are most interesting in your project. (You do not need to copy your safety forms onto this wiki page.)</p>
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<section id="cta" style="background-image:url('https://static.igem.org/mediawiki/2015/6/65/Unitn_pics_cta_safety.jpg');">
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<header>
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<h2><strong> Safety </strong></h2>
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<p> Work hard, work safe </p>
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</header>
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</section>
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<!-- BEGIN iGEM_Content_Section -->
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<article id="main">
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<section class="wrapper style1 container special" style="margin-top:3em;">
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<div class="row">
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<div class="4u 12u(narrower)">
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<section>
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<header class="wow zoomIn" style="visibility:hidden;">
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<div href="#" class="rotate-box square-icon"><span class="rotate-box-icon"><i class="faa flaticon-bacteria"></i></span></div>
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</header>
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<p>We have reviewed our organisms and parts against the <a class="i_linker" href="https://2015.igem.org/Safety/WhiteList" target="_blank">White List</a>. All the parts and bacterial strains that we used are in the <span class="i_enph">Risk Group 1</span></p>
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</section>
  
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</div>
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<div class="4u 12u(narrower)">
  
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<h4>Safe Project Design</h4>
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<section>
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<header class="wow zoomIn" style="visibility:hidden; -moz-animation-delay: 0.2s; -webkit-animation-delay: 0.2s; animation-delay: 0.2s;">
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<div href="#" class="rotate-box square-icon zoomIn"><span class="rotate-box-icon"><i class="faa flaticon-checkmark11"></i></span></div>
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</header>
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<p>We have submitted to iGEM headquarters <span class="i_enph">“About Our Lab”</span> and <span class="i_enph">“About Our Project”</span> questionnaires by the June 26<sup>th</sup> deadline</p>
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</section>
  
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<p>Does your project include any safety features? Have you made certain decisions about the design to reduce risks? Write about them here! For example:</p>
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</div>
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<div class="4u 12u(narrower)">
  
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<ul>
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<section>
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<li>Choosing a non-pathogenic chassis</li>
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<header class="wow zoomIn" style="visibility:hidden; -moz-animation-delay: 0.5s; -webkit-animation-delay: 0.5s; animation-delay: 0.5s;">  
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<li>Choosing parts that will not harm humans / animals / plants</li>
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<div href="#" class="rotate-box square-icon zoomIn"><span class="rotate-box-icon"><i class="faa flaticon-verified7"></i></span></div>  
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<li>Substituting safer materials for dangerous materials in a proof-of-concept experiment</li>
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</header>
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<li>Including an "induced lethality" or "kill-switch" device</li>
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<p>We have submitted to iGEM headquarters the <span class="i_enph">“Final Safety Form”</span> by the August 28<sup>th</sup> deadline</p>
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</ul>
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</section>
  
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<h4>Safe Lab Work</h4>
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</div>
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</div>
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</section>
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<!-- One -->
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<section class="wrapper style4 container wow fadeInRight" style="margin-top:1em;">
  
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<p>What safety procedures do you use every day in the lab? Did you perform any unusual experiments, or face any unusual safety issues? Write about them here!</p>
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<div class="content">
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<section>
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<header>
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<h3 class="wow fadeInDown" style="visibility:hidden;">About Safety</h3>
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</header>
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 +
<p>We, the UNITN iGEM Team want to <span class="i_enph">follow high standards of safe and responsible biological engineering</span>. Before entering the lab we attended mandatory on-line courses regarding lab safety with final tests. In particular we focused on the following topics:</p>
 +
<ul class="customlist checklist">
 +
<li>General considerations on risks and prevention</li>
 +
<li>Electrical Hazard</li>
 +
<li>Chemical Hazard</li>
 +
<li>Biological Hazard</li>
 +
<li>Compressed gasses and cryogenic liquids</li>
 +
</ul>
 +
<p>We also attended the “Health and safety at the workplace” course, which is mandatory for all students and workers at the University of Trento.</p>
 +
</section>
 +
</div>
  
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<h4>Safe Shipment</h4>
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</section>
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<section class="wrapper style4 container wow fadeInLeft" style="margin-top:1em; -moz-animation-duration: 0.5s; -webkit-animation-duration: 0.5s; animation-duration: 0.5s;">
  
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<p>Did you face any safety problems in sending your DNA parts to the Registry? How did you solve those problems?</p>
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<!-- Content -->
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<div class="content">
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<section>
 +
<!--<a href="#" class="image featured"><img src="images/pic04.jpg" alt="" /></a>-->
 +
<header>
 +
<h3 class="wow fadeInDown" style="visibility:hidden;">Safe Project Design</h3>
 +
</header>
 +
<!-- TODO -->
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 +
<div class="row">
 +
<div class="6u 12u(narrower)">
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<section>
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<header>
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<p class="question wow fadeInDown" style="visibility:hidden; -moz-animation-delay: 0.5s; -webkit-animation-delay: 0.5s; animation-delay: 0.5s;">Who will use your product?</p>
 +
</header>
 +
<p style="margin-top:1em">Our solar microbial fuel cell is designed to be <span class="i_enph">used by the public</span>. We envision that our Solar pMFC will be implemented to allow people with <span class="i_enph">no specific skills to operate it</span>.</p>
 +
</section>
 +
</div>
 +
<div class="6u 12u(narrower)">
 +
<section>
 +
<header>
 +
<p class="question wow fadeInDown" style="visibility:hidden; -moz-animation-delay: 0.5s; -webkit-animation-delay: 0.5s; animation-delay: 0.5s;">Where will your product be used?</p>
 +
</header>
 +
<p style="margin-top:1em">Our idea is to have a device that can be installed <span class="i_enph">in your house</span> and mimics a photovoltaic system. What is needed is just a good exposition to light, e.g. placing it on top of a roof.</p>
 +
</section>
 +
</div>
 +
</div>
 +
 +
<header>
 +
<p class="question wow fadeInDown" style="visibility:hidden; -moz-animation-delay: 0.5s; -webkit-animation-delay: 0.5s; animation-delay: 0.5s;">What happens when it's all used up? Will it be sterilized, discarded, or recycled?</p>
 +
</header>
 +
 +
<a class="fancybox wow zoomIn" rel="group" href="https://static.igem.org/mediawiki/2015/e/e7/Unitn_pics_safety_0.jpg" style="margin-top:0; display:block; height:320px; background-image:url('https://static.igem.org/mediawiki/2015/e/e7/Unitn_pics_safety_0.jpg'); background-size:cover;  background-position-y: 62%;"></a>
 +
 +
<p style="margin-top:1em; margin-bottom:0.5em;">Our MFC requires a growth media, our engineered bacteria and sunlight. In the laboratory we  tested the system with <span class="i_enph">LB</span> or <span class="i_enph">M9</span>, however we envision that our system will be optimized so that the bacteria can <span class="i_enph">use organic waste from the house</span> (i.e. kitchen waste containing sugars or other carbon sources). To minimize the amount of contaminated media we were thinking to implement the <span class="i_enph italic">Solar pMFC</span> with a circulating system equipped with a filter that allows to remove the bacteria  from the media, while accumulating the bacteria in the filter. In this way the <span class="i_enph">decontaminated  media could be disposed directly in the plumbing system of the house</span>.</p>
 +
   
 +
<p>The exhausted bacteria  accumulated in the filter could be routinely removed and discarded properly (not in the  environment). Fresh bacteria could be added placing a new batch of bacteria in the system. We  imagine a capsule, similar to espresso-capsule, that could be placed easily in the system when  necessary. The life-span of the bacteria today it is a limit, although our Proteorhodopsin expressing bacteria should live significant longer, thus reducing the need of replacing them often.</p>
 +
 
 +
<header>
 +
<p class="question wow fadeInDown" style="visibility:hidden; -moz-animation-delay: 0.5s; -webkit-animation-delay: 0.5s; animation-delay: 0.5s;">Is it safer, cheaper, or better than other technologies that do the same thing?</p>
 +
</header>
 +
 +
<p style="margin-top:1em">Our Solar pMFC is better than existing technologies because:</p>
 +
<ul class="customlist arrowed" style="margin-top:0; margin-left:2em;">
 +
<li>We know exactly the bacteria that are inside and generate electricity, thus reducing the biological hazard. We believe that using <span class="i_enph">well characterized and controlled</span> (i.e inducible) <span class="i_enph">bacteria provides a better-working and safer device</span>. This is an advantage respect to other MFCs.</li>
 +
<li>It uses sun light to be powered, thus <span class="i_enph">reducing the consumption of electricity</span> to operate it.</li>
 +
<li>Thank to our circulating/filter system to decontaminate the media, we will <span class="i_enph">reduce the amount of dirty waste</span> to be disposed, thus making it better and safer for the environment.</li>
 +
<li>It is better than photovoltaic systems on the market that are based on silicon. Silicon it is not a  problem itself, however the production of <span class="i_enph">silicon photovoltaic systems requires toxic chemicals</span>  (silicon tetrachloride, Sulphur Hexafluoride, heavy metals such cadmium and lead). Also, the  disposal of large amount of silicon panels will become a problem one day for the environment.</li>
 +
<li>Our pMFC will be <span class="i_enph">cheap</span> and thus be competitive on the market. The production cost of one solar pMFC to be used in the laboratory is around  <span class="i_enph great-quantity">250 <span>$</span></span>. This cost covers the expenses of the material
 +
and the work to build a cell containing 1.5 L of media. <span class="i_enph">This cost should be decreased at least by half</span> if the MFC is built industrially.  Alternatively in a no distant future we will be able to print MFCs at home with a 3D printer.<br />We estimated that today the cost of operation for 48 hours is:<br />
 +
<span class="i_enph great-quantity">15.00 <span>$</span></span> of media. <span class="i_enph italic">This cost should become zero</span> if our bacterial strain become able to use alternative carbon sources, i.e. kitchen waste.<br />
 +
<span class="i_enph great-quantity">0.02 <span>$</span></span> for arabinose.<br />
 +
<span class="i_enph great-quantity">0.00 <span>$</span></span> for sunlight<br />
 +
Therefore <span class="i_enph italic">the operating cost should become close to zero</span>.
 +
</li>
 +
</ul>
 +
 
 +
</section>
 +
</div>
 +
</section>
 +
 +
 +
<section class="wrapper style4 container wow fadeInRight" style="margin-top:1em;">
  
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<!-- Content -->
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<div class="content">
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<section>
 +
<header>
 +
<h3 class="wow fadeInDown" style="visibility:hidden;">Working Safely... with Parts</h3>
 +
</header>
 +
<!-- TODO -->
 +
 +
 +
<div class="row">
 +
<div class="4u 12u(narrower) centered">
 +
<a class="fancybox" rel="group" href="https://static.igem.org/mediawiki/2015/b/be/Unitn_pics_safety_1.jpg"><img src="https://static.igem.org/mediawiki/2015/b/be/Unitn_pics_safety_1.jpg" alt="" style="width:100%; max-width:500px;"/></a>
 +
</div>
 +
<div class="8u 12u(narrower)">
 +
<p>All the parts we used throughout the project are <span class="i_enph">NOT toxic or dangerous for  humans</span> or the environment. We neither worked with proteins toxic themselves, nor we used enzymes that synthesize toxic molecules. In addition to basic parts from the registry (i.e. RBS, promoters, β-carotene, etc), we also used two genes from the SAR86 uncultured bacteria. These genes are proteorhodopsin, which was extracted from the Registry (BBa_K773002) and blh (15,15’-β-carotene dioxygenase), which was synthesized by Genescript. There is no associated risk level with the uncultured bacteria SAR86, however we are using only DNA sequences that encodes for genes, which carry distinct functions that are no harmful to humans, or the environment. We have clarified this issue with iGEM Safety Office.</p>
 +
</div></div>
 +
 +
<p>We did <span class="i_enph">NOT use any virulence factors</span> in our organisms.<br />
 +
The parts we used are to be considered safe as single parts and also as combined. Environmental dispersion of those parts would <span class="i_enph">NOT represent a biological hazard or danger</span>.</p>
 +
</div>
 +
 +
<header>
 +
<h3 class="wow fadeInDown" style="visibility:hidden;">Working Safely... with Organisms</h3>
 +
</header>
 +
<p>All the bacterial strains used are from Risk Group 1. We used the following strains:</p>
 +
<ul class="customlist checklist">
 +
<li> One Shot® TOP10 Chemically Competent E. coli</li>
 +
<li> K12 - NEB 10-beta E. coli</li>
 +
<li> K12 - JM109 E. coli</li>
 +
<li> BL21 - NEB Express E. coli</li>
 +
</ul>
 +
</section>
 +
</div>
 +
</section>
 +
 +
 +
<section class="wrapper style4 container wow fadeInLeft" style="margin-top:1em;">
  
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<h3 class="wow fadeInDown" style="visibility:hidden;">Personal Protective Equipment (PPE)</h3>
 +
</header>
 +
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<div class="row">
 +
<div class="4u 12u(narrower)">
 +
<a class="fancybox" rel="group" href="https://static.igem.org/mediawiki/2015/d/dd/Unitn_pics_safety_2.jpg"><img src="https://static.igem.org/mediawiki/2015/5/5d/Unitn_pics_safety_2_thumb.jpg" alt="" style="width:100%; max-width:500px;"/></a>
 +
</div>
 +
 +
<div class="8u 12u(narrower)">
 +
<p>Last but non least we adopted the <span class="i_enph">appropriate PPE during the lab work</span>.  We always used lab coats, nitrile gloves and goggles.  We safely operated with biological material under the laminar flow cabinet <b>BioAir SAFEMATE Series Class II 1.8</b>.</p>
 +
<p>We used chemicals that are associated with hazard (ethidium bromide, azide,  organic solvents) under the chemical hood <b>Fume Hood MOD. ASEM 120EN New - Class 0</b>.</p>
 +
<p>We carefully revised all the <b>Material Safety Datasheet</b> of each compound that was used prior to start an experiment.</p> 
 +
</div>
 +
</div>
 +
</section>
 +
</div>
 +
</section>
 +
 
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Latest revision as of 01:14, 17 September 2015

Safety

Work hard, work safe

We have reviewed our organisms and parts against the White List. All the parts and bacterial strains that we used are in the Risk Group 1

We have submitted to iGEM headquarters “About Our Lab” and “About Our Project” questionnaires by the June 26th deadline

We have submitted to iGEM headquarters the “Final Safety Form” by the August 28th deadline

We, the UNITN iGEM Team want to follow high standards of safe and responsible biological engineering. Before entering the lab we attended mandatory on-line courses regarding lab safety with final tests. In particular we focused on the following topics:

  • General considerations on risks and prevention
  • Electrical Hazard
  • Chemical Hazard
  • Biological Hazard
  • Compressed gasses and cryogenic liquids

We also attended the “Health and safety at the workplace” course, which is mandatory for all students and workers at the University of Trento.

Our solar microbial fuel cell is designed to be used by the public. We envision that our Solar pMFC will be implemented to allow people with no specific skills to operate it.

Our idea is to have a device that can be installed in your house and mimics a photovoltaic system. What is needed is just a good exposition to light, e.g. placing it on top of a roof.

Our MFC requires a growth media, our engineered bacteria and sunlight. In the laboratory we tested the system with LB or M9, however we envision that our system will be optimized so that the bacteria can use organic waste from the house (i.e. kitchen waste containing sugars or other carbon sources). To minimize the amount of contaminated media we were thinking to implement the Solar pMFC with a circulating system equipped with a filter that allows to remove the bacteria from the media, while accumulating the bacteria in the filter. In this way the decontaminated media could be disposed directly in the plumbing system of the house.

The exhausted bacteria accumulated in the filter could be routinely removed and discarded properly (not in the environment). Fresh bacteria could be added placing a new batch of bacteria in the system. We imagine a capsule, similar to espresso-capsule, that could be placed easily in the system when necessary. The life-span of the bacteria today it is a limit, although our Proteorhodopsin expressing bacteria should live significant longer, thus reducing the need of replacing them often.

Our Solar pMFC is better than existing technologies because:

  • We know exactly the bacteria that are inside and generate electricity, thus reducing the biological hazard. We believe that using well characterized and controlled (i.e inducible) bacteria provides a better-working and safer device. This is an advantage respect to other MFCs.
  • It uses sun light to be powered, thus reducing the consumption of electricity to operate it.
  • Thank to our circulating/filter system to decontaminate the media, we will reduce the amount of dirty waste to be disposed, thus making it better and safer for the environment.
  • It is better than photovoltaic systems on the market that are based on silicon. Silicon it is not a problem itself, however the production of silicon photovoltaic systems requires toxic chemicals (silicon tetrachloride, Sulphur Hexafluoride, heavy metals such cadmium and lead). Also, the disposal of large amount of silicon panels will become a problem one day for the environment.
  • Our pMFC will be cheap and thus be competitive on the market. The production cost of one solar pMFC to be used in the laboratory is around 250 $. This cost covers the expenses of the material and the work to build a cell containing 1.5 L of media. This cost should be decreased at least by half if the MFC is built industrially. Alternatively in a no distant future we will be able to print MFCs at home with a 3D printer.
    We estimated that today the cost of operation for 48 hours is:
    15.00 $ of media. This cost should become zero if our bacterial strain become able to use alternative carbon sources, i.e. kitchen waste.
    0.02 $ for arabinose.
    0.00 $ for sunlight
    Therefore the operating cost should become close to zero.

All the parts we used throughout the project are NOT toxic or dangerous for humans or the environment. We neither worked with proteins toxic themselves, nor we used enzymes that synthesize toxic molecules. In addition to basic parts from the registry (i.e. RBS, promoters, β-carotene, etc), we also used two genes from the SAR86 uncultured bacteria. These genes are proteorhodopsin, which was extracted from the Registry (BBa_K773002) and blh (15,15’-β-carotene dioxygenase), which was synthesized by Genescript. There is no associated risk level with the uncultured bacteria SAR86, however we are using only DNA sequences that encodes for genes, which carry distinct functions that are no harmful to humans, or the environment. We have clarified this issue with iGEM Safety Office.

We did NOT use any virulence factors in our organisms.
The parts we used are to be considered safe as single parts and also as combined. Environmental dispersion of those parts would NOT represent a biological hazard or danger.

All the bacterial strains used are from Risk Group 1. We used the following strains:

  • One Shot® TOP10 Chemically Competent E. coli
  • K12 - NEB 10-beta E. coli
  • K12 - JM109 E. coli
  • BL21 - NEB Express E. coli

Last but non least we adopted the appropriate PPE during the lab work. We always used lab coats, nitrile gloves and goggles. We safely operated with biological material under the laminar flow cabinet BioAir SAFEMATE Series Class II 1.8.

We used chemicals that are associated with hazard (ethidium bromide, azide, organic solvents) under the chemical hood Fume Hood MOD. ASEM 120EN New - Class 0.

We carefully revised all the Material Safety Datasheet of each compound that was used prior to start an experiment.